MD1-60 is presented as 96 x 1 ml conditions in a deep-well block.

Transcription

1 MIDAS TM HT-96 MD1 60 MIDAS TM : a Modern Intelligent Dynamic Alternative Screen - A revolutionary 96 condition crystallization screen based on alternative polymeric precipitants 1. MD1-60 is presented as 96 x 1 ml conditions in a deep-well block. Features of MIDAS: Ideal for soluble protein, protein/protein complexes, protein-nucleic acid complexes and sensitive macromolecular complexes. Narrow range of ph and salt concentrations centered on physiological values. Every condition contains at least one alternative polymeric precipitant. Designed to complement PEG and salt-based screens. Compatible with liquid-handling robots. There are many alternatives to PEGs and have recently been described as being useful for macromolecular crystallogenesis. Alternative polymers (Figure 2) such as the Jeffamine polyetheramines, pentaerythritol propoxylate and pentareythritol, polyvinyl pyrrolidone, polypropylene glycol, polyvinyl alcohol and polyacrylate have so far only sporadically been introduced into standard crystallization screens. Introduction MIDAS is a 96 condition crystallization screen based on alternative polymeric precipitants. Devised and tested (Figure 1) in the Laboratory of Dr. Clemens Grimm et al of Würzburg University in Germany. MIDAS is a revolutionary crystallization screen that has moved away from the reliance on polyethylene glycols (PEGs) as the main precipitant (only 3 conditions in MIDAS contain a PEG). MIDAS systematically searches for crystallization conditions with alternative polymeric precipitants. MIDAS entails a relatively narrow range of ph and salt concentrations centred on physiological values to increase its suitability for sensitive macromolecular complexes, while every condition contains at least one alternative polymeric precipitant. PEG Alternatives: For decades PEGs or their monomethyl ethers (PEG MMEs), have dominated crystallization screens. Out of 8289 entries scanned in the PDB, almost half of the crystallization conditions contained a PEG component and most commercial screens available today contain PEGs. However, the success rate of PEGs might be influenced due to their widespread dominance in crystallization screens. 1 Figure 1. Examples of protein crystals grown using MIDAS. (A) Lysozyme crystals obtained in 35% Sokalan HP 56, (B) Xylanase crystals obtained in 20% Jeffamine M2070, (C) Crystals of the cytokine receptor ligand complex obtained in 45% pentaerythritol propoxylate (5/4 PO/OH), (D) Crystals of streptavidin core obtained in 5% polyacrylate 2100, sodium salt, (E) Histone tail recognizing MBT repeats in 35% polyacrylate 2100, sodium salt), (F) Lysozyme crystals in 30% Sokalan CP 42,(G) spliceosomal assembly complex (SAC) 7 obtained in 6% polyvinyl pyrrolidone, (H) Crystals of spliceosomal assembly complex (SAC) 9 obtained in 25% Sokalan CP 42.

2 Formulation Notes: MIDAS reagents are formulated using ultrapure water (>18.0 MΩ) and are sterile-filtered using 0.22 µm filters. No preservatives are added. Final ph may vary from that specified on the datasheet. Molecular Dimensions will be happy to discuss the precise formulation of individual reagents. Individual reagents and stock solutions for optimization are available from Molecular Dimensions. Enquiries regarding MIDAS formulation, interpretation of results or optimization strategies are welcome. Please e- mail, fax or phone your query to Molecular Dimensions. Contact and product details can be found at Manufacturer s safety data sheets are available to download from our website. Pentaerythritol ethoxylate. Jeffamine ED2003 Pentaerythritol propoxylate Jeffamine SD2001 References : 1. Grimm, C., Chari, A., Reuter, K. & Fischer, U. (2010). Acta Cryst. D66, Poly(acrylic acid-co-maleic) acid polyvinylpyrrolidone M-type Jeffamines* Vinylpyrrolidone/vinylimidazole Copolymer polypropylene glycol polyvinyl alcohol Figure 2. Examples of alternative precipitants used in MIDAS TM, *R1 = H for EO or CH3 for PO. The PO/EO molar ratio is 29/6 for Jeffamine M2005, 10/31 for Jeffamine M2070 and 9/1 for Jeffamine M600. MIDAS is manufactured and distributed under an exclusive license with Dr. C. Grimm & Prof. Dr. U. Fischer. Limited Use and Restrictions: Products sold by Molecular Dimensions Ltd. or its affiliates or authorized distributors and information relating to same are intended for research use only in crystal growth and optimization of crystal growth following use of the product by the purchaser and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, including resale or use in manufacture. The license to use MIDAS TM specifically excludes any rights to use the product information for the manufacture of the product or derivatives thereof, or distribute, transfer, or otherwise provide access to such information to any third party for any purpose or use. 2

3 MIDAS HT-96 Conditions A1-D12 MD1 60 Well # Conc. Salt Conc. Buffer ph Conc. Precipitant A1 0.1 M HEPES % v/v Polypropylene glycol % v/v Dimethyl sulfoxide A2 0.1 M MES % w/v Polyvinylpyrrolidone A3 0.1 M HEPES % w/v Poly(acrylic acid sodium salt) 2100 A4 14 % v/v Poly(acrylic acid-co -maleic acid) solution A5 0.5 M Ammonium phosphate monobasic 12.5 % w/v Poly(acrylic acid sodium salt) 2100 A6 0.1 M Tris % v/v Poly(acrylic acid-co -maleic acid) solution A7 10 % v/v Polypropylene glycol 400 A8 5 % w/v Poly(acrylic acid sodium salt) 2100 A9 0.1 M MES % v/v Pentaerythritol propoxylate (5/4 PO/OH) A M Sodium sulfate 24 % w/v Polyvinylpyrrolidone A M Calcium chloride dihydrate 0.1 M HEPES % v/v Pentaerythritol ethoxylate (15/4 EO/OH) A M Potassium/sodium phosphate % v/v Polypropylene glycol 400 B1 0.2 M Sodium chloride 0.1 M MES % v/v Jeffamine D % v/v Jeffamine M-2005 B2 0.2 M Sodium thiocyanate 0.1 M HEPES % v/v Pentaerythritol propoxylate (5/4 PO/OH) B3 0.2 M Potassium acetate 0.1 M HEPES % w/v Polyvinyl alcohol 10 % v/v Jeffamine T-403 B4 0.2 M Sodium chloride 0.1 M MES % v/v Pentaerythritol propoxylate (5/4 PO/OH) B5 0.1 M HEPES % w/v Polyvinyl alcohol 10 % v/v 1-Propanol B6 0.1 M Lithium sulfate 0.1 M HEPES % w/v Polyvinylpyrrolidone B7 0.2 M Imidazole % v/v Polypropylene glycol 400 B M Lithium sulfate 0.1 M HEPES % v/v Poly(acrylic acid-co -maleic acid) solution 3 % v/v Pentaerythritol ethoxylate (3/4 EO/OH) B9 0.1 M Sodium chloride 0.1 M Tris % v/v Jeffamine SD-2001 B10 30 % v/v Jeffamine M % v/v Dimethyl sulfoxide B11 20 % v/v Polypropylene glycol % v/v 1-Propanol B M HEPES % v/v Poly(acrylic acid-co -maleic acid) solution C1 15 % v/v Jeffamine ED-2003 C2 0.2 M Sodium chloride 0.1 M MES % v/v Jeffamine ED-2003 C3 0.1 M Sodium malonate dibasic monohydrate 0.1 M MES % v/v Jeffamine SD-2001 C4 0.2 M Sodium chloride 0.1 M MES % v/v Pentaerythritol propoxylate (5/4 PO/OH) C5 0.2 M Magnesium chloride hexahydrate 35 % v/v Pentaerythritol ethoxylate (3/4 EO/OH) C6 40 % v/v Pentaerythritol propoxylate (5/4 PO/OH) 15 % v/v Ethanol C7 0.1 M Tris % v/v Pentaerythritol propoxylate (5/4 PO/OH) C8 0.2 M Sodium chloride 0.1 M Tris % w/v Polyvinylpyrrolidone 10 % w/v PEG 4000 C9 0.1 M Sodium chloride 25 % v/v Pentaerythritol propoxylate (5/4 PO/OH) 10 % v/v Dimethyl sulfoxide C M Ammonium sulfate 0.1 M HEPES % w/v Poly(acrylic acid sodium salt) 2100 C M Magnesium formate dihydrate 0.1 M Tris % v/v Pentaerythritol ethoxylate (15/4 EO/OH) C M Potassium acetate 24 % v/v Poly(acrylic acid-co -maleic acid) solution D1 0.1 M Tris % v/v Polypropylene glycol 400 D2 0.1 M HEPES % v/v Pentaerythritol ethoxylate (15/4 EO/OH) 6 % w/v Polyvinylpyrrolidone D3 45 % v/v Polypropylene glycol 400 D4 10 % v/v Pentaerythritol ethoxylate (3/4 EO/OH) 10 % v/v 1-Butanol D5 0.1 M HEPES % w/v Poly(acrylic acid sodium salt) % v/v Jeffamine SD-2001 D6 0.1 M HEPES % w/v Polyvinylpyrrolidone D7 0.1 M HEPES % v/v Jeffamine ED-2003 D8 0.1 M Tris % v/v Glycerol ethoxylate 10 % v/v Tetrahydrofuran D9 0.2 M Imidazole % v/v Jeffamine D-2000 D M Potassium chloride 0.1 M HEPES % v/v Jeffamine SD-2001 D M Sodium chloride 30 % v/v Polypropylene glycol 400 D12 20 % v/v Jeffamine SD % v/v 1-Propanol 3

4 MIDAS HT-96 Conditions E1- H12 MD1 60 Well # Conc. Salt Conc. Buffer ph Conc. Precipitant E1 0.2 M Lithium sulfate 0.1 M Tris % v/v Jeffamine T-403 E2 0.2 M Potassium acetate 35 % v/v Pentaerythritol propoxylate (5/4 PO/OH) E3 0.2 M Potassium chloride 0.1 M Glycine % v/v Pentaerythritol ethoxylate (15/4 EO/OH) E4 0.2 M Sodium thiocyanate 0.1 M HEPES % v/v Pentaerythritol propoxylate (5/4 PO/OH) E5 0.2 M Potassium chloride 0.1 M HEPES % v/v Jeffamine T % v/v Jeffamine ED-2003 E6 0.2 M Potassium acetate 0.1 M MES % v/v Pentaerythritol ethoxylate (15/4 EO/OH) 3 % v/v Jeffamine T-403 E7 0.1 M Sodium malonate dibasic monohydrate 0.1 M HEPES % w/v Poly(acrylic acid sodium salt) 2100 E8 10 % v/v Jeffamine D % v/v Jeffamine M-2005 E9 0.1 M Lithium sulfate 0.1 M Tris % v/v Jeffamine ED-2003 E M Tris % v/v Jeffamine T % v/v Jeffamine ED-2003 E M Lithium sulfate 0.1 M HEPES % w/v Poly(acrylic acid sodium salt) 2100 E M Magnesium chloride hexahydrate 0.1 M HEPES % w/v Poly(acrylic acid sodium salt) 2100 F1 0.1 M HEPES % v/v Jeffamine D-2000 F2 0.5 M Sodium chloride 0.1 M Tris % w/v Poly(acrylic acid sodium salt) 2100 F3 0.1 M Potassium/sodium phosphate % v/v Jeffamine ED % v/v Jeffamine SD-2001 F4 0.2 M Sodium chloride 0.1 M BICINE % w/v Poly(acrylic acid sodium salt) 2100 F5 0.2 M Sodium malonate dibasic monohydrate 0.1 M MES % v/v Jeffamine D-2000 F6 0.2 M Potassium chloride 0.1 M Tris % v/v Jeffamine M-2070 F7 20 % v/v Jeffamine M % v/v Dimethyl sulfoxide F8 0.2 M Magnesium chloride hexahydrate 0.1 M MES % v/v Pentaerythritol propoxylate (17/8 PO/OH) F9 0.1 M Tris % w/v Poly(acrylic acid sodium salt) 5100 F M HEPES % v/v Polyethyleneimine F M Ammonium formate 0.1 M HEPES % v/v SOKALAN CP 7 F M Sodium sulfate 0.1 M Tris % w/v SOKALAN HP 56 G1 0.1 M Potassium chloride 0.1 M HEPES % v/v SOKALAN CP 7 G2 0.3 M Ammonium formate 0.1 M HEPES % w/v SOKALAN CP 5 G3 40 % v/v Glycerol ethoxylate G4 0.1 M Tris % v/v Glycerol ethoxylate G5 55 % v/v Polypropylene glycol 400 G6 0.2 M Lithium citrate tribasic tetrahydrate 35 % v/v Glycerol ethoxylate G7 0.2 M Ammonium acetate 0.1 M MES % v/v Glycerol ethoxylate G8 0.1 M Tris % w/v SOKALAN CP 42 5 % v/v Methanol G9 0.1 M Tris % w/v SOKALAN CP % v/v Tetrahydrofuran G M Lithium acetate dihydrate 0.1 M Bis-Tris % w/v SOKALAN CP 42 G M HEPES % v/v Jeffamine M-2005 G M Bis-Tris % w/v SOKALAN CP 5 H1 0.1 M Bis-Tris % w/v SOKALAN CP 42 H2 35 % v/v Jeffamine D-2000 H3 0.1 M Tris % v/v Glycerol ethoxylate 3 % v/v Polyethyleneimine H4 0.2 M Ammonium chloride 0.1 M HEPES % v/v Glycerol ethoxylate H5 0.1 M Tris % w/v SOKALAN CP 42 H6 0.1 M MES % w/v Poly(acrylic acid sodium salt) 5100 H7 0.2 M Potassium citrate tribasic monohydrate 15 % w/v SOKALAN CP 42 H8 0.1 M Tris % w/v SOKALAN CP 42 H9 0.2 M Ammonium acetate 0.1 M HEPES % w/v SOKALAN HP 56 H M Tris % w/v SOKALAN CP 5 H M Ammonium formate 10 % w/v Polyvinylpyrrolidone 20 % w/v PEG 4000 H M Tris % w/v Polyvinylpyrrolidone 25 % w/v PEG 5000 MME 4

5 Abbreviations: BICINE: 2-(Bis(2-hydroxyethyl)amino)acetic acid, Bis Tris; Bis-(2-hydroxyethyl)iminotris(hydroxymethyl)methane, HEPES; 2-(4-(2-Hydroxyethyl)-1-piperazinyl)ethanesulfonic Acid, MES; 2-(Nmorpholino)ethanesulfonic acid, PEG; Polyethylene glycol, Tris; 2-Amino-2-(hydroxymethyl)propane-1,3-diol. N.B. J. Polyvinylpyrolidone K15 is now called Polyvinylpyrolidine. The conditions shown on this datasheet may differ from those shown on previous versions of the datasheets (only applicable to MIDAS batches ) due to the discontinuation of raw material supply for the following: Glascol W13, Sokalan CP12S and Sokalan HP66K. If you require further advice regarding the changes to these conditions please contact us at Sokalan are water-soluble polymers based on acrylic acid, maleic acid, vinylpyrrolidone, vinylimmadazole and/or hydrophobic monomers. Manufacturer s safety data sheets are available from our website or by scanning the QR code here: Re-Ordering details: Catalogue Pack size Catalogue Code MIDAS 96 x 10 ml MD1-59 MIDAS HT x 1 ml MD1-60 MIDAS Green Screen 96 x 10 ml MD1-83 MIDAS HT-96 Green Screen 96 x 1 ml MD1-84 MIDAS OptiMax Kit* 24 x 10 ml MD1-62 Single Reagents MIDAS single reagents 100 ml MDSR-59-tube number MIDAS HT96 single reagents 100 ml MDSR-60-well number For MIDAS stock solutions please visit the Optimization section on our website. *MIDAS OptiMax contains all the individual stock reagents for MIDAS. 5